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2005 Fiscal Year Final Research Report Summary

Development and applications of a protozoan parasite vector using Toxoplasma gondii to express heterologous genes

Research Project

Project/Area Number 14360172
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Basic veterinary science/Basic zootechnical science
Research InstitutionObihiro University of Agriculture and Veterinary Medicine

Principal Investigator

XUAN Xuenan  Obihiro University, Professor, 原虫病研究センター, 教授 (10292096)

Co-Investigator(Kenkyū-buntansha) YOKOYAMA Naoaki  Obihiro University, Assoc.Prof., 原虫病研究センター, 助教授 (80301802)
INOUE Noboru  Obihiro University, Assoc.Prof., 原虫病研究センター, 助教授 (10271751)
TANAKA Tetsuya  Hokkaido University, Assist.Prof., 農学部, 助手 (00322842)
IWATA Akira  Nippon Institute for Biological science, Senior Researcher, 主任研究員 (70193745)
TAKASHIMA Yasuhiro  Gifu University, Assoc.Prof., 応用生物科学部, 助教授 (20333552)
Project Period (FY) 2002 – 2005
KeywordsT.gondii / N.canunim / C.parvum / NcSRS2 / CpP23 / Recombinant vaccine / Vector vaccine / Protection
Research Abstract

An improved method for the construction of recombinant Toxoplasma gondii expressing heterologous genes was developed using pyrimethamine-resistant DHFR-TS and green fluorescent protein (GFP) genes as double selection markers. This improved method allows the construction of recombinant T.gondii expressing foreign genes with great case and speed. Because the foreign genes were placed between the DHFR-TS and GFP genes, all of the drug-resistant and fluorescent clones were confirmed to be expressed foreign genes. This method was employed to construct recombinant T.gondii expressing the Neospora caninum major antigen SRS2 gene (Tg/NcSRS2) and the Cryptosporidium parvum major antigen P23 gene (Tg/CpP23). The expression levels of NcSRS2 and CpP23 in recombinant Tg/NcSRS2 and Tg/CpP23 were similar to those in N.caninum and C.parvum, respectively. The molecular weight and antigenic property of recombinant NcSRS2 and CpP23 were similar to those of native NaSRS2 and CpP23, respectively. Mice immunized with recombinant Tg/NcSRS2 were partially protected from N.caninum challenge infection. On the other hand, the mice immunized with recombinant Tg/CpP23 produced specific neutralizing antibodies against C.parvum. In addition, a bioactive molecule, such as mouse interferon gamma was successfully expressed by T.gondii vector. These results indicate that the T.gondii vector may provide a new tool not only for the production of recombinant vaccines against protozoan parasite infections in animals, but also for the production of bioactive molecules.

  • Research Products

    (13 results)

All 2005 2004 2003 2002

All Journal Article (12 results) Book (1 results)

  • [Journal Article] Identification and characterization of cross-reactive antigens from Neospora caninum and Toxoplasma gondii.2005

    • Author(s)
      Liao et al.
    • Journal Title

      Parasitology 130巻

      Pages: 481-488

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Expression of P23 of Cryptosporidium parvum in Toxoplasma gondii and evaluation of its protective effects.2005

    • Author(s)
      Shirafuji et al.
    • Journal Title

      J. Parasitol. 91巻

      Pages: 476-479

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Identification and characterization of cross-reactive antigens from Neospora caninum and Toxoplasma gondii.2005

    • Author(s)
      Liao, M., Xuan, X., Shirafuji, H., Fukumoto, S., Hirata, H., Suzuki, H., Fujisaki, K.
    • Journal Title

      Parasitol. 130

      Pages: 481-488

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Expression of P23 of Cryptosporidium parvum in Toxoplasma gondii and evaluation of its protective effects.2005

    • Author(s)
      Shirafuji, H., Xuan, X., Kimata, I., Takashima, Y., Shinya, F., Otsuka, H., Nagsdawa, H., Suzuki, H.
    • Journal Title

      J.Parasitol. 91

      Pages: 476-479

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Natural IgM antibodies in sera from various animals but not the cat kill Toxoplasma gondii by activating complement classical pathway. Parasitol.2004

    • Author(s)
      Kaneko et al.
    • Journal Title

      Parasitology 128巻

      Pages: 123-129

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Natural IgM antibodies in sera from various animals but not the cat kill Toxoplasma gondii by activating complement classical pathway.2004

    • Author(s)
      Kaneko, Y., Takashima, Y., Xuan, X., Igarashi, I., Nagasawa, H., Mikami, T., Otsuka, H.
    • Journal Title

      Parasitol. 128

      Pages: 123-129

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Characterization of Toxoplasma gondii engineered to express mouse interferon-gamma.2003

    • Author(s)
      Nishikawa et al.
    • Journal Title

      Int. J. Parasitol. 33巻

      Pages: 1525-1535

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Characterization of Toxoplasma gondii engineered to express mouse interferon-gamma.2003

    • Author(s)
      Nishikawa, Y., Xuan, X., Makala, L., Vielemeyer, O., Joiner, K.A., Nagasawa, H.
    • Journal Title

      Int.J.Parasitol 33

      Pages: 1525-1535

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Neospora caninum NcSRS2 is a transmembrane protein that contains a glycosylphosphatidylinositol anchor in insect cells.2002

    • Author(s)
      Nishikawa et al.
    • Journal Title

      Vet. Parasitol. 109巻

      Pages: 191-201

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Recombinant feline herpesvirus type 1 expressing Toxoplasma gondii ROP2 antigen inducible protective immunity in cats. 2002.2002

    • Author(s)
      Mishima et al.
    • Journal Title

      Parasitol. Res. 88巻

      Pages: 144-149

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Neospora caninum NcSRS2 is a transmembrance protein that contains a glycosylphosphatidylinositol anchor in insect cells.2002

    • Author(s)
      Nishikawa, Y., Tragoolpua, K., Makala, L., Xuan, X., Nagasawa, H.
    • Journal Title

      Vet.Parasitol. 109

      Pages: 191-201

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Recombinant feline herpesvirus type 1 expressing Toxoplasma gondii ROP2 antigen inducible protective immunity in cats.2002

    • Author(s)
      Mishima, M., Xuan, X., Yokoyama, N., Igarashi, I., Fujisaki, K., Nagasawa, H., Mikami, T.
    • Journal Title

      Parasitol.Res. 88

      Pages: 144-149

    • Description
      「研究成果報告書概要(欧文)」より
  • [Book] 獣医微生物学2003

    • Author(s)
      玄学南(見上彪監修)
    • Total Pages
      307-312
    • Publisher
      文永堂出版
    • Description
      「研究成果報告書概要(和文)」より

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Published: 2007-12-13  

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